Sequence polymorphism can produce serious artefacts in real-time PCR assays: hard lessons from Pacific oysters.

ABSTRACT:

Background

Since it was first described in the mid-1990s, quantitative real time PCR (Q-PCR) has been widely used in many fields of biomedical research and molecular diagnostics. This method is routinely used to validate whole transcriptome analyses such as DNA microarrays, suppressive subtractive hybridization (SSH) or differential display techniques such as cDNA-AFLP (Amplification Fragment Length Polymorphism). Despite efforts to optimize the methodology, misleading results are still possible, even when standard optimization approaches are followed.

Results

As part of a larger project aimed at elucidating transcriptome-level responses of Pacific oysters (Crassostrea gigas) to various environmental stressors, we used microarrays and cDNA-AFLP to identify Expressed Sequence

SUBMITTER: Taris N 

PROVIDER: S-EPMC2424065 | biostudies-literature | 2008 May

REPOSITORIES: biostudies-literature